Rapid Molecular Strategy for Orbivirus Detection and Characterization

Palacios, Gustavo; Cowled, Christopher; Bussetti, Ana Valeria; Savji, Nazir; Weir, Richard; Wick, Ivan; Rosa, Amélia Travassos da; Calisher, Charles H.; Tesh, Robert B.; Boyle, David; Lipkin, W. Ian

doi:10.1128/jcm.00337-11

Cited by 19 publications

(13 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Ross River virus was detectable in both 100 and 1000 pools, highlighting the sensitivity of the method, and detection was not affected by centrifugation of samples prior to extraction. These results encourage testing for a wider range of arboviruses using group‐reactive primers (for example Eshoo et al., ; Kuno, ; Palacios et al., ). Due to the multiplexing capabilities of NGS, many large pools of vectors could be simultaneously screened for a panel of arboviruses, thereby improving the efficiency of virus screening.…”

Section: Discussionmentioning

confidence: 83%

Effective mosquito and arbovirus surveillance using metabarcoding

Batovska

Lynch

Cogan

et al. 2017

Molecular Ecology Resources

View full text Add to dashboard Cite

Effective vector and arbovirus surveillance requires timely and accurate screening techniques that can be easily upscaled. Next‐generation sequencing (NGS) is a high‐throughput technology that has the potential to modernize vector surveillance. When combined with DNA barcoding, it is termed ‘metabarcoding.’ The aim of our study was to establish a metabarcoding protocol to characterize pools of mosquitoes and screen them for virus. Pools contained 100 morphologically identified individuals, including one Ross River virus (RRV) infected mosquito, with three species present at different proportions: 1, 5, 94%. Nucleic acid extracted from both crude homogenate and supernatant was used to amplify a 269‐bp section of the mitochondrial cytochrome c oxidase subunit I (COI) locus. Additionally, a 67‐bp region of the RRV E2 gene was amplified from synthesized cDNA to screen for RRV. Amplicon sequencing was performed using an Illumina MiSeq, and bioinformatic analysis was performed using a DNA barcode database of Victorian mosquitoes. Metabarcoding successfully detected all mosquito species and RRV in every positive sample tested. The limits of species detection were also examined by screening a pool of 1000 individuals, successfully identifying the species and RRV from a single mosquito. The primers used for amplification, number of PCR cycles and total number of individuals present all have effects on the quantification of species in mixed bulk samples. Based on the results, a number of recommendations for future metabarcoding studies are presented. Overall, metabarcoding shows great promise for providing a new alternative approach to screening large insect surveillance trap catches.

show abstract

Section: Discussionmentioning

confidence: 83%

Effective mosquito and arbovirus surveillance using metabarcoding

Batovska

Lynch

Cogan

et al. 2017

Molecular Ecology Resources

View full text Add to dashboard Cite

show abstract

“…Of particular interest are those based on broad spectrum RT-PCR approaches. There are several conventional or real-time pan-generic PCR approaches for the identification of flaviviruses (Moureau et al, 2007), orbiviruses (Palacios et al, 2011) and bunyaviruses, particularly pan-nairovirus, pan-orthobunyavirus and pan-phlebovirus PCRs (Lambert and Lanciotti, 2009). Full-length characterizations may be realized using virus sequence-independent methodologies described for dsRNA including the single primer amplification technique (SPAT, Attoui et al, 2000) or the SPAT-derived full length amplification of complementary cDNA (FLAC, Maan et al, 2007), or for ssRNA such as that combining a FLAC/SMART (Switch Mechanism At the 5′ end of RNA Template) methodology (Attoui et al, 2007).…”

Section: Do Endogenous Viruses Play a Modulating Role In Infection Ofmentioning

confidence: 99%

Endogenous tick viruses and modulation of tick-borne pathogen growth

Bell‐Sakyi¹,

Attoui²

2013

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

Ticks transmit a wide range of viral, bacterial and protozoan pathogens, many of which can establish persistent infections of lifelong duration in the vector tick and in some cases are transmitted transovarially to the next generation. In addition many ixodid and argasid tick cell lines and, by inference the parent ticks from which they were derived, harbor endogenous viruses (ETV) of which almost nothing is known. In general, low level persistent infections with viral pathogens (arboviruses) are not known to have a deleterious effect on tick survival and fitness, suggesting that they can strike a balance with the tick innate immune response. This tolerance of arbovirus infection may be modulated by the permanent presence of ETV in the host cell. In mosquito cells, temporary or permanent silencing of the genes of an endogenous virus by RNA interference can result in changes in replication rate of a co-infecting arbovirus. We propose that tick cell lines offer a useful model system for in vitro investigation of the modulatory effect of ETV on superinfecting pathogen survival and replication in ticks, using the molecular manipulation techniques applied to insect cells.

show abstract

“…Orbiviruses are known to infect multiple animal species (Attoui et al, 2011). Traditionally orbivirus isolates were classified based on their serological properties (Attoui et al, 2005;Belaganahalli et al, 2011Belaganahalli et al, , 2012Palacios et al, 2011). The development of next generation sequencing technologies have repeatedly demonstrated their utility in identifying non-cultivable viruses and also in characterizing existing virus isolates that cannot be identified by more traditional laboratory methods (Victoria et al, 2008).…”

mentioning

confidence: 99%

“…The development of next generation sequencing technologies have repeatedly demonstrated their utility in identifying non-cultivable viruses and also in characterizing existing virus isolates that cannot be identified by more traditional laboratory methods (Victoria et al, 2008). Many studies that included all known orbiviruses have demonstrated that phylogenetic analysis and genetic relatedness can be used to classify uncharacterized orbiviruses (Belaganahalli et al, 2011(Belaganahalli et al, , 2012Palacios et al, 2011). Moreover, the orbiviruses transmitted by a common arthropod vector (i.e.…”

mentioning

confidence: 99%

A novel mosquito-borne Orbivirus species found in South-east Asia

Kapoor

Tesh

Duraisamy

et al. 2013

Journal of General Virology

Self Cite

View full text Add to dashboard Cite

The genus Orbivirus of the family Reoviridae includes a genetically diverse group of dsRNA arthropod-borne viruses that infect a wide variety of animal species. Here, we report the complete genome and phylogenetic analysis of a novel orbivirus (IAn-66411 or Sathuvachari virus, SVIV) isolated in 1963 from starlings (Brahminy myna) collected in Vellore, Tamil Nadu, India. Comparative genetic analysis of the SVIV polymerase (VP1 protein), core protein (VP3) and outer core protein (VP7) confirmed that SVIV is most closely related to the mosquito-borne orbiviruses, but that it is equally divergent from all known species. Therefore, SVIV should be tentatively considered as the prototype of a novel mosquito-associated Orbivirus species. These findings will aid in the development of molecular reagents that can identify genetically similar orbiviruses and help elucidate their geographical distribution, epidemiology, species tropism and possible disease association.

show abstract

Rapid Molecular Strategy for Orbivirus Detection and Characterization

Cited by 19 publications

References 12 publications

Effective mosquito and arbovirus surveillance using metabarcoding

Effective mosquito and arbovirus surveillance using metabarcoding

Endogenous tick viruses and modulation of tick-borne pathogen growth

A novel mosquito-borne Orbivirus species found in South-east Asia

Contact Info

Product

Resources

About